Mass Spectrometry of RNA-Binding Proteins during Liquid–Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures

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dc.citation.firstpage10659
dc.citation.issueNumber19
dc.citation.journalTitleJournal of the American Chemical Society
dc.citation.lastpage10668
dc.citation.volumeNumber145
dc.contributor.authorSahin, Cagla
dc.contributor.authorMotso, Aikaterini
dc.contributor.authorGu, Xinyu
dc.contributor.authorFeyrer, Hannes
dc.contributor.authorLama, Dilraj
dc.contributor.authorArndt, Tina
dc.contributor.authorRising, Anna
dc.contributor.authorGese, Genis Valentin
dc.contributor.authorHällberg, B. Martin
dc.contributor.authorMarklund, Erik. G.
dc.contributor.authorSchafer, Nicholas P.
dc.contributor.authorPetzold, Katja
dc.contributor.authorTeilum, Kaare
dc.contributor.authorWolynes, Peter G.
dc.contributor.authorLandreh, Michael
dc.contributor.orgCenter for Theoretical Biological Physics
dc.date.accessioned2023-07-21T16:13:56Z
dc.date.available2023-07-21T16:13:56Z
dc.date.issued2023
dc.description.abstractLiquid–liquid phase separation (LLPS) of heterogeneous ribonucleoproteins (hnRNPs) drives the formation of membraneless organelles, but structural information about their assembled states is still lacking. Here, we address this challenge through a combination of protein engineering, native ion mobility mass spectrometry, and molecular dynamics simulations. We used an LLPS-compatible spider silk domain and pH changes to control the self-assembly of the hnRNPs FUS, TDP-43, and hCPEB3, which are implicated in neurodegeneration, cancer, and memory storage. By releasing the proteins inside the mass spectrometer from their native assemblies, we could monitor conformational changes associated with liquid–liquid phase separation. We find that FUS monomers undergo an unfolded-to-globular transition, whereas TDP-43 oligomerizes into partially disordered dimers and trimers. hCPEB3, on the other hand, remains fully disordered with a preference for fibrillar aggregation over LLPS. The divergent assembly mechanisms revealed by ion mobility mass spectrometry of soluble protein species that exist under LLPS conditions suggest structurally distinct complexes inside liquid droplets that may impact RNA processing and translation depending on biological context.
dc.identifier.citationSahin, Cagla, Motso, Aikaterini, Gu, Xinyu, et al.. "Mass Spectrometry of RNA-Binding Proteins during Liquid–Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures." <i>Journal of the American Chemical Society,</i> 145, no. 19 (2023) American Chemical Society: 10659-10668. https://doi.org/10.1021/jacs.3c00932.
dc.identifier.digitaljacs-3c00932
dc.identifier.doihttps://doi.org/10.1021/jacs.3c00932
dc.identifier.urihttps://hdl.handle.net/1911/115002
dc.language.isoeng
dc.publisherAmerican Chemical Society
dc.rightsExcept where otherwise noted, this work is licensed under a Creative Commons Attribution (CC BY) license.  Permission to reuse, publish, or reproduce the work beyond the terms of the license or beyond the bounds of Fair Use or other exemptions to copyright law must be obtained from the copyright holder.
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.titleMass Spectrometry of RNA-Binding Proteins during Liquid–Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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